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. Author manuscript; available in PMC: 2008 Jan 1.
Published in final edited form as: Anal Biochem. 2006 Nov 3;360(1):151–153. doi: 10.1016/j.ab.2006.10.019

Figure 2.

Figure 2

TLE as a method to assay Asp/Glu-Adt. A. Phosphorimage of a TLC plate developed in Solvent A. The origin is not shown and is approximately 2.25 inches below the lowest spot. B. TLE plate electroeluted for 40 minutes at 100 volts in Solvent B. The solid line denotes the site of sample loading (origin); the directions of the cathode (+) and the anode (-) are also labeled. In both A and B, each lane was spotted at the origin with 50 nCi of the relevant 14C-labelled amino acid, as labeled. Lane 1-Asp; Lane 2-Asn; Lane 3-1:1 mixture of Asp:Asn; Lane 4-Glu; Lane 5-Gln; Lane 6-1:1 ratio of Glu:Gln. C. An assay of Asp/Glu-Adt using TLE. Asp/Glu-Adt was assayed as described in the text. Lanes 1-3 represent amino acid controls (Lane 1-Asp, Lane 2-Asn, Lane 3-1:1 ratio of Asp:Asn). Lane 4 shows the results from the no enzyme control where the Asp-tRNAAsn remains unreacted. Lane 5 shows the results from the Asp/Glu-Adt assay where approximately 50% of the Asp-tRNAAsn has been converted to Asn-tRNAAsn.